| 牛乳铁蛋白基因N-lobe的cDNA克隆及毕赤酵母表达载体的构建 |
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| 引用本文： | 林松涛,张居农,王亮,史芳芳,李卫华,张钰,吕自力,王安江.牛乳铁蛋白基因N-lobe的cDNA克隆及毕赤酵母表达载体的构建[J].家畜生态,2008(5):13-18. |
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| 作者姓名： | 林松涛 张居农 王亮 史芳芳 李卫华 张钰 吕自力 王安江 |
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| 作者单位： | 中国科学院新疆理化技术研究所;石河子大学动物科技学院;新疆金牛生物有限公司; |
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| 基金项目： | 新疆维吾尔自治区高新技术项目(编号:200611104) |
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| 摘 要： | 以牛乳腺组织总RAN为模板,经RT-PCR得到牛乳铁蛋白基因的N-lobe片段并将其克隆到pGM-T克隆载体上,最后将其连入表达载体pPICZαA中,构建牛乳铁蛋白基因N-lobe的酵母表达载体。结果表明：克隆片段大小为1028bp,与Gen Bank中登录的序列的相应部位相比,所获牛乳铁蛋白N-lobe cDNA序列的同源性为99.7%;重组表达质粒经酶切和PCR鉴定构建正确,为酵母表达乳铁蛋白基因N-lobe奠定了基础。
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| 关 键 词： | 乳铁蛋白 N-lobe 表达载体 |
cDNA Cloning of Bovine Lactoferrin N-lobe Gene and Construction of Pichia Pastoris Expression Vector |
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| LIN Song-tao,ZHANG Ju-nong,WANG Liang,SHI Fang-fang,LI Wei-huaZHANG Yu,LV Zi-li,WANG An-jiang.cDNA Cloning of Bovine Lactoferrin N-lobe Gene and Construction of Pichia Pastoris Expression Vector[J].Ecology of Domestic Animal,2008(5):13-18. |
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| Authors: | LIN Song-tao ZHANG Ju-nong WANG Liang SHI Fang-fang LI Wei-huaZHANG Yu LV Zi-li WANG An-jiang |
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| Institution: | LIN Song-tao1,2,ZHANG Ju-nong2,WANG Liang1,3,SHI Fang-fang3,LI Wei-hua3ZHANG Yu3,LV Zi-li3,WANG An-jiang3(1.Xinjiang Technical Institute of Physics , Chemistry,CAS,Urumqi 830011,2.College of Animal Science , Technology,Shihezi Univercity,Shihezi,Xinjiang 832003,3.Xinjiang Gold-Cattle Biology Incorporation,Urumqi 830026,China) |
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| Abstract: | Using the total RAN from bovine mammary grand as a template to construct Pichia pastoris Expression Vector of bovine Lactoferrin N-lobe Gene. the N- lobe gene of bovine lactoferrin have been amplified with RT-PCR method. The obtained DNA fragment was then cloned into pGM-T vector. Pos - itive recombinant plasmid was digested with EcoRl and XbaI and the N-lobe fragment was reco- rered. The Pichia pastoris expression Vector was constructed by inserting the N-lobe fragment into vector of pPICZα A The sequencing results of recombinant cloning vector demonstrated the obtained N-lobe gene has 1028bp and 99.7 % homology with it＇s gene in GenBank. The sequencing results of recombinant Expression vector proved by restriction enzymes and sequencing that our construction is correct. It provides bases for Expression of N-lobe in Pichia pastoris. |
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| Keywords: | Lactoferrin N-lobe Base Sequence Expression Vector |
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